The goal of this Program Project is to study basic mechanisms of induced immune unresponsiveness in an attempt to understand: (1) the pathogenesis of autoimmune disease and (2) the potential for the development of innovative new immunotherapeutics for the treatment or prevention of autoimmune disease. The first Project studies the lymphocyte multifunctional calcium-dependent calmodulin protein kinase (CaM kinase) which, in preliminary studies using transfection with a "constitutive mutant," has been demonstrated to block IL-2 transcription. The objective of this first Project is to further study the possible role CaM kinase plays in T cell anergy. The second Project utilizes the murine model of experimental allergic encephalomyelitis (EAE) as a model of autoimmune disease in which several different modes of immunotherapy have been developed which seem to result in a form of unresponsiveness resembling T cell clonal anergy. The goals of the second Project interrelate with those of the first Project through the attempt to isolate "anergy genes" which might be potential targets of immune intervention and/or dysregulation in autoimmunity. The third Project relates directly to potential immunotherapy of a murine model of autoimmunity, IDDM, in the NOD mouse. This Project will assess the potential regulatory effect of TNF-alpha on disease pathogenesis and as a potential immunotherapeutic to block development of disease. The final Project focuses on novel combinations of transgene-positive mice to probe regulation of islet cell-reactive T cells and/or the production of autoantibodies reactive with islet cells. We believe these four projects attempt to explore fundamental issues of immune unresponsiveness. They additionally relate induced immune unresponsiveness to potential immunotherapy of autoimmune disease and allow the study of dysregulation of immune unresponsiveness which may allow the induction of autoimmunity.